Frog Probiotics

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Welcome to frogprobiotics.org! This site is designed to highlight our project to find effective probiotics for Malagasy amphibians in order to develop a proactive conservation response for the impending arrival of the deadly amphibian pathogen, Batrachochytrium dendrobatidis, in Madagascar. Check out the Madagascar tab to learn more. We provide information for donors who want to support our effort to protect frogs in Madagascar. For more information about this project and how to donate please contact: Reid Harris and Molly Bletz.

Saturday, February 15, 2014

I can’t believe it is getting to the end of my time here in
Madagascar.7 weeks seemed so long on
the first day but it went by with a blink of the eye. I spent our final days sampling frogs at the
Vohimana reserve and the Mitsinjo Forest.In my mind, I thought I had seen enough frogs but when we caught some
Boophis albilabris at Vohimana and five Spinomantis aglavai in Mitsinjo forest,
it was quite a treat!

Sampling for this trip was completed on Feb 10th
and we have successfully collected over 500 samples for probiotics from over 90
species of frogs. I couldn’t be happier with the efforts of this trip!

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Spinomantis aglavai

The last couple of days I have been having meetings in Tana
with representatives from the Department of Forestry, the Cellule d’Urgence
Chytride, Durrell, MNP, Conservation International, Ambatovy, Veterninary
Sciences and Animal Biology Department of the University of Tana to discuss our
collaborative initiative to work toward finding effective probiotics for Malagasy
amphibians.On the first day, we
discussed the background about Bd, the killer amphibian fungus, and I also
presented on our proactive plan for conservation of amphibians using
probiotics.We showed the group here the
preliminary results from the laboratory work in the USA – showing that we have 7
bacterial isolates that can inhibit Bd by at least 95%. The afternoon was
filled with discussions about the next steps and how we can work together to
rapidly identify effective probiotics.It was an exhausting but successful day!﻿﻿﻿﻿﻿﻿

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On day 2, we spent the day at the University, and I
demonstrated for the stakeholders here the methods of the activities we have
been doing in the USA, including bacteria culturing and DNA extraction, and
presented a tutorial on the methods of qPCR. We also toured some of the labs to
determine how we can build capacity here in Madagascar for isolating probiotic
bacteria.

Boophis luteus

On the third and final day we were joined by another US researcher
Jonathan Kolby who will be working with the Cellule d’Urgence Chytride to
sample Bd in Madagascar. We finalized our discussion of probiotics and
discussed that if and when Bd arrived what are the best ways to rapidly respond and
conserve Malagasy frogs. These meetings with the government were most productive and encouraging!

On Monday, I will be on a plane back to the States.A few days ago I would have said I was ready
to return from the grueling field work…But already I feel the call of the
jungle and frogs that is beckoning me to stay.I can’t wait to return on our next trip to continue sampling and
building capacity here in Madagascar to save the frogs!

Wednesday, February 5, 2014

It was great to be back with the frogs after our taxi
brousse adventure.Our next adventure
took us to the forest around Fierenana, just north of Moramanga. The road to
get there was an adventure in itself.Victor our driver did a fantastic job maneuvering around the potholes,
steering clear of the landslides, and crossing bridges just wide enough for the
suspension of the land cruiser. We managed to get stuck only twice! But when we
got stuck we were really stuck.

Rice fields of Fierenana

Five-six hours of bumping around in the car and pushing to
car out of the mud twice we arrived in Fierenana.We got there much later than expected so we
stayed the night in a local family's house. After talking with the local
guide we found out that the site with the Mantellamilotympanum we wanted to visit was
at least a 4 hr hike.We planned to
start around 4:30 AM to get to the site mid-morning but of course in Malagasy
time the porters arrived around 5:30.Eventually we were on our way to the home of Mantellamilotympanum.The area around Fierenana is known for its
rice production and the seemingly endless expanse of rice patties spoke truth
to that. Not surprisingly, the trek began with a long expanse of rice
patties.It was a balancing act to walk
of the small dirt mounds dividing the rice patties. Most of the journey there
was traversing rice patties and dirt roads on deforested slopes with some
patches of remaining forest.After about
16 km we reached what the guide told us was the sight.Devin looked at his GPS and with a puzzled
looked explained to the guide that the coordinates of the site were still a few
km away.We mustered up the energy to
continue and the trail entered a patch of forest which was much more enjoyable
than the rice patties.

Mantella milotympanum

After another hour or two of walking we came to a clearing
with a small house and Devin said we were within a km of the site.The porters and we were exhausted from our 23
km trek so we decided to set up camp in the grassy field by the house (with the
approval of the owner of course).Luckily the rain let up when we were setting up camp. It was so nice to
finally get our packs of and rest before sampling at night.

Boophis rappiodes

We travelled to a nearby river for sampling and there were
tons of Boophisrappiodes calling along with other species.We sampled the frogs that night and got
rested up for our early morning Mantella searching.We started around 6 and headed to a marsh that
looked promising from the night before. As we arrived you could already here them
calling and within minutes we caught the first one. They are such a cute
frog!I also stumbled upon a Dyscophusguineti which was unexpected and cool. We ended the morning with
about 20 Mantella and 4 Dyscophus which was a success for both
Karina and I.

The next morning it was already time to hike out so we
suited up packs and all and began our trek out.We had walked so far in that it was actually closer to hike about to
another village called Ambohinydri. The trek was a series of rice fields, then a hike over a forested slope, then through another rice field, and up and over another
forested slope.It seemed as though
every valley had been taken over by rice. By the end the lower half of my legs
were coated in mud from the muddy paths and miss-steps in the rice patties. By
2 our 20+ km trek out was complete and we were one our way back to Andasibe.

The trip was a success and we now have samples for potential
probiotics from the endangered Mantellamilotympanum!

Friday, January 31, 2014

Our last few days in Ranomafana went very well. We travelled to the Vohiparara circuit with another frog group doing taxonomy work. The night was filled with mating frenzies of Guibemantis liber, Blommersia blommersae, Aglyptodactylus madagascariensis and many new species of tree frogs! The leeches were relentless as always.

We were scheduled to leave ValBio on the morning of the 19th with our photographer friends from Antoetra. Their vehicle arrived at the station, and they jumped out to meet us with some perplexed looks. Evidently the driver had decided our gear could not go on top of the car and also couldn’t block the back window. We relentlessly tried to fit all the gear in the vehicle. We stuffed as much gear as possible in the back below the window and figured we would put the rest on our laps, but then the driver said "no," that was not OK either. The driver claimed it was a police violation and despite our efforts it was a no go.

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Male Aglyptodactylus madagascariensis

Slightly panicked because I was unsure of how we would get back to Tana for our departure the next day to Ambohintantely, we decided our only option was a taxi brousse. The morning direct from Ranomafana to Tana had already left, and it was Sunday so travelling is always difficult. We talked with the ValBio guard, and he agree to flag down a local brousse to Fianaransoa - the closest major town. After two hours one finally past and jammed ourselves into an already over filled bus. Che and I more or less had to share one seat. As we traveled the curvy road at full speed, there was no way to not squish your neighbor. I was surrounded by a few children and their mothers and some teenage boys just in front of us and a chicken or two clucked from below the seat. Two of the kids were feeling queasy with the curvy road. Luckily there were some small plastic bags on board. The mother was siting backwards behind the driver’s seat. Without a glace over her shoulder she tossed the baggy of throw up out the window and there was a load SMACK! She had released the baggy just as a car came wising by in the other direction. The taxi brousse came to a quick halt. We were stopped for about a half an hour as the woman discussed who knows what with the driver. They made her clean the vehicle with the only thing she had which was a small sweatshirt. After a while I guess they came to some kind of consensus and we all piled back into the taxi and continued on our way to Fianar. We finally made it after bumping up and down and side to side for 3 hours.
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Boophis guibei

The only taxi brousse to Tana was the overnight bus that left at 7 PM. We had some waiting to do before our second adventure. The time passed relatively quickly and soon enough we were sitting on the taxi waiting to depart. The rain had begun and it was already leaking through the sliding door, and the driver’s window was nothing more than a sheet of torn plastic. The driver duct taped the one door… hopefully just for the leaking rain and we were not depending on that duct tape to keep the door closed. Fortunately we ended up departing early because the taxi was full.

The music blared loudly and we were on our to be 12 hour ride to Tana. We tried to sleep but the pot holes and sharp turns made it nearly impossible. For some reason Taxi brousses are stopped at just about every checkpoint by the police. We would come to a screeching halt and the police would shine there lights through the rows of bodies and we would be on our way. We stopped for dinner around 9 in Ambositra and enjoy an extremely large bowl of soup. Within about 5 minutes it seemed like we were on the taxi brousse holding our breath that the car would start.

Some hours later my seat neighbor was kicked out of the taxi, and we were joined by 2 policemen was AK 47s… not exactly what I had in mind for the rest of our journey. In about an hour they left and we gained a few more passengers making it a bit crowded in our row. In Antsirabe, our taxi brousse was officially out of commission and we had to switch to a different vehicle. Luckily there was one waiting for us, making the 4 am switch quite smooth. This new taxi brousse was outfitted with much more comfortable seats so getting a little shut-eye was somewhat possible. However, this driver was a very aggressive driver whipping around every turn at at least 80 km/h if not more. Around 5, we started picking up people along the road and soon there were 6 people squeezed in our row meant for 4 people and one person sitting backwards against the front passenger seat. The last 2 hours I was squished unable to move in any direction. But by 7 we had finally reached Tana and grabbed a taxi to the hotel where we had hoped to spend the night. We had survived the taxi brousse and made it to Tana before our planned departure at 8 am for Ambohintantely… so I would call our adventure a success.

Saturday, January 18, 2014

When a boa 2 meters long falls from a tree at your feet, the
night resounds with the chirps, trills, and clicks of over 100 frogs species,
fist-sized snails glide over the forest floor and insects of any shape, color
and size crawl and buzz around, you know you are in the rainforest, one of the
most magnificent habitats on earth (in my opinion). In the rainforest of Madagascar you are also greeted by blood thirsty terrestrial leeches, that somehow no matter what you wear find a way in. Serge and I spent 2
nights in Vaoharanana a site in Ranomafana National Park.The hike in had some unforgiving ups and
downs but the dense canopy of the rainforest sheltered us from the afternoon
sun which was nice. We all made it to the site in one piece and the cook had
already started lunch.After lunch we
started searching for frogs in the leaf litter and along the stream.

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Mantidactylus cowani "small"

The stream was teaming with Mantidactylus
cowani “small,” which was really cool!We
found 8 different species the first day sampling. Around 4 PM the clouds rolled
in and the sky began to rumble with thunder.Serge whispered to me as we waited for dinner “I bet we will have over
50 frogs tonight.”The rain began to
pour down and didn’t let up until 8 PM but we sampled anyway.Within seconds I was drenched from head to
toe.It was almost pointless to wear a
raincoat. The leeches were ever so relentless with the rainy night. Every
minute or so I would feel there suckers inching their way up my leg or
neck.I had even duct taped my pant leg
down to my skin to prevent them from crawling in but somehow they found a way
in.We ended the night with 9 new
species and many new ones from our last trip in August, including a Platypeli
grandis – a truly amazing frog.Its toe
pads are so big, almost too big for its body, sort of like when a dog’s paws
seem too big for it when it’s a puppy.We processed the frogs with the help of the guide.Serge has mastered a one-handed frog hold, which is great for saving the gloves we use for sampling.We went to sleep in our tents, which were
luckily safely set up under tin roof structures to keep us happy and dry.

Platypelis grandis

By the next morning the rain had past and the sun danced through the trees and across the stream, giving it a magical sparkle.The cook prepared us some coffee and
breakfast- i.e. a huge portion of rice and some fried zebu.In the morning we collected along the small
tributary streams and the forest floor upstream from the camp.We found 6 new species in addition to some of
the ones found the previous day. We had
lunch and the processed the frogs in the early afternoon. Serge and I have
become rather efficient at sampling the frogs so it goes pretty quickly. The
rest of the afternoon we rested and organized for our second and final sampling
night at Vato. Packs of lemurs noisily rumbled through the forest making quite
a ruckus, but we were unable to seen them.The thunder and rain rolled in earlier around 2 PM, but by 6:30 PM it
was more of a light mist which was preferable to the torrential rain the
previous night.This was the night were
a huge boa fell from a tree just beside me as I searched for a calling Guibemantis
liber. When it fell I jumped and charged full force away from it stumbling over
logs and getting tangled in lianas and bamboo grass, not knowing was it
was.I creep back over to find a 2 meter
boa now resting on the forest floor.With my heart still racing a bit, I decided to venture to another
section of the forest near a small stream away from the big snake.

Gephyromantis tschenki

Along the small stream, the Boophis quasibohemi were calling
from every direction.I saw one male
proudly grasping the back of his new mate, and probably soon they would deposit
and fertilize some eggs dangled above the stream. The night ended with 4 new
species.

We had sampled 108 individuals of 30 different species by
the end of our 2 days at Vato, and the terrestrial leeches had sampled us many times (33 for me!). We also
saw many metamorphs hopping around the forest floor and tadpoles darting around
the small tributaries and heard many more species calling from high up in the
canopy.The frogs here seemed happy and
healthy!

Friday, January 10, 2014

It was great to finally make it into the countryside of Madagascar. We, including Karina, Richard, Serge, our student from University of Tana, Devin from Association Mitsinjo, Brian a photographer from Cal Academy and me, were greeted with the rolling hills of green, interspersed with grey rock outcrops and dotted with small villages here and there. The drive from Ambositra to Antoetra was short compared our 6 hour drive the Tana the previous day. In Antoetra we were met with the voices of smiling children shouting Vazaha, which means foreigner in Malagasy. With relative ease (Thanks to Serge and Devin) we arranged for a guide and a cook for our trek to find Mantella cowani in the hills surrounding Antoetra. Our gear required 11 porters for the six of us and soon enough our small army took off on the trail toward Farimazava. After about 1.5 hours of hiking we arrived at our camp location. The owner of a house in the village there graciously opened their home to us, and we all got to escape the rain and cool nights in a room in the attic. The owner of the house had a welcoming speech for us and we all had a sip of some home brewed rum. I was anxious to sample the first frogs of the trip so we went out to a small stream to find frogs and came back with over 20 frogs for the night including Boophis microtympanum which was a unique find.

The late night sampling made the 4:30 AM wake up for searching for Mantella a bit challenging, but the scenic views of waterfalls and the pinkish orange sunrise soon broke my tiredness. After about a 45 minute walk we arrived at the Farimazava site. The site looked somewhat deforested and I was a bit skeptical at first about whether there would be any frogs. But the locals know there stuff. Soon we found a rocky, fast flowing stream and just up the stream a 100 m or so we began to heard the chirps of the Mantella! The site was filled with M. baroni and a few putative baroni-cowani hybrids which was great for Karina’s work. We left empty handed for cowani but we would look again tomorrow at another site. The day felt so long after the early wake up but rewarding because we got to sample the first wild mantella of the trip. During the afternoon I showed the children around the house the frog ID book and they were mesmerized by the photos and would tell me the local names for the different herps in the book. We took the night off to rest and gear up for the early morning trek in search of the Sahona mana as the locals call M. cowani. We wandered the hillsides of a historic locality of cowani and ears and eyes open, but unfortunately no cowani. We did however, find some a few frogs that were either Mantidactylus lugubris or Mantidactylus cowan which was a new species for our project.

Set on finding Mantella cowani we decided to hike out of the village, Holoma Ambany Lalana, and search a site south of Antoetra called Soamazaka. We spend the night in Antoetra and headed to the site just after sunrise. Ranary, our driver was able to get us very close to the site which was much appreciated after our uphill hike the day before. We had two local guides with us and within seconds of arrival one was pointing at the side of the rocky stream and we all scrambled over. It was our first Mantella cowani! All of our faces lit up in excitement at the sight of our target species. Within 2 hours we had found 9 cowani and a few other species like Boophis goudoti and Mantidactylus curtus. Processing went quickly and smoothly. Our time in Antoetra was indeed a success!

We all stayed in Ambositra for the night, before our planned departure to Isalo the next day. Devin and Brian were headed back to Andasibe, and Karina, Richard, Serge and I left early expecting the long travel to Ranohira (near Isalo). Along the way I glanced over our permit localities and to my surprise Isalo was not listed as a sampling locality! In a slight panic, I showed the permit to Serge and asked him what we should do. Then we contacted our collaborator Faly Rabemananjara in Tana to see what he thought we should do. He said we had two options: drive to Isalo and talked with the forestry office there to see if they would let us sample in the Isalo area or go to our next site Ranomafana. We had a decision to make: continue on the long 10 hours car ride to Ranohira and hope the forestry office would grant us permission to sample or go to Ranomafana and do more extensive sampling of the forests there.

We decided to go to Ranomafana and then see if we could contact the office in Isalo by phone. A day past and we still could not find a number to call. We decided that it would be more productive and logistically easier for us to stay in Ranomafana. This park has one of the most diverse amphibian assemblages of Madagascar, with over 125 species. Mantella expectata, one our target species at Isalo will have to wait to be sampled on the next trip. We head into the forest in Ranomafana tomorrow to begin sampling in the park to determine the prevalence of Bd and collect probiotic samples. The rain is pouring down today and will hopefully bring out the frogs!

Wednesday, January 8, 2014

Molly is proceeding to the Isalo area today to sample potential probiotics from Mantella expectata, as well as other species. This species is very rare and will definitely need protection if Bd arrives in this area. Isalo is in the southern part of Madagascar and differs from the rainforest habitats of Andasibe and Ranomafana that we have sampled so far. It is generally drier and characterized by oases where frogs congregate during the wet season.

Meanwhile, the JMU crew is continuing to isolate and test bacterial isolates collected from Malagasy frogs during the August and September trip for effective anti-Bd probiotics.